Project 2: Synthetic Lethal Therapy for HPV-Negative Head and Neck Cancer

项目 2：HPV 阴性头颈癌的合成致死疗法

• 批准号: 10913240
• 负责人: BARBARA BURTNESS
• 金额: $ 7.41万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-22 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10913240
• 关键词: Acceleration; Affect; Aneuploidy; Apoptosis; Biological Markers; CASP3 gene; CDC2 gene; CDKN2A gene; CHEK1 gene; Cell Cycle; Cell Cycle Arrest; Cell Cycle Checkpoint; Cell Line; Cell Survival; Cells; Centrosome; Characteristics; Cisplatin; Clinic; Clinical; Clinical Trials; Complement; Cyclin-Dependent Kinases; DNA Damage; DNA Repair; DNA Sequence Alteration; Data; Defect; Dependence; Doctor of Philosophy; Dose; Drug Combinations; Drug Targeting; Effectiveness; Enrollment; Evaluation; Exhibits; Freezing; Future; G1/S Checkpoint Pathway; Generations; Genes; Genomics; Goals; HPV-negative head and neck cancer; Head and Neck Cancer; Head and Neck Neoplasms; Head and Neck Squamous Cell Carcinoma; Human; Human Papillomavirus; Inflammatory; Lead; Malignant Neoplasms; Measures; Mitosis; Mitotic; Mitotic spindle; Modeling; Mus; Mutate; Mutation; Normal Cell; Normal tissue morphology; Operative Surgical Procedures; PLK1 gene; Patient Selection; Patients; Pharmaceutical Preparations; Pharmacodynamics; Phase; Phenotype; Phosphorylation; Phosphotransferases; Ploidies; Population; Protein Dephosphorylation; Protein-Serine-Threonine Kinases; Recommendation; Resectable; Resistance; Serine; Signal Transduction; TP53 gene; Testing; Toxic effect; Translating; Tumor Suppressor Proteins; Work; Xenograft Model; Xenograft procedure; anti-cancer; aurora kinase A; biomarker identification; cancer cell; checkpoint inhibition; cohort; cytotoxic; cytotoxicity; effective therapy; exome sequencing; experimental study; gain of function; genomic biomarker; inhibitor; kinase inhibitor; neoantigens; novel; patient biomarkers; patient derived xenograft model; pharmacologic; pre-clinical; predictive marker; preservation; repaired; replication stress; response; response biomarker; single cell sequencing; synergism; targeted agent; therapy resistant; translational impact; tumor

Summary HPV-negative head and neck squamous cell carcinomas (HNSCC) typically lose G1/S cell cycle checkpoints, with most tumors having mutations in TP53, and many also mutating other tumor suppressors such as CDKN2A. Such tumors become dependent on checkpoints associated with G2/M to repair DNA damage arising from replication stress and other genomic insults. This dependency suggests a tumor-selective vulnerability to synthetic lethal strategies controlling progress through G2/M. In extensive preliminary data, we show that AZD1775/adavosertib, an inhibitor of the G2/M checkpoint kinase WEE1, potently sensitizes TP53mut HNSCC cell lines to inhibition of Aurora A kinase (AURKA). WEE1 induces an inhibitory Y15-phosphorylation of CDK1, blocking M-phase entry and thus blunting the cytotoxic effects AURKA inhibition. WEE1 inhibition abrogates this arrest, accelerating mitotic entry for cells bearing highly disruptive spindle abnormalities and other defects arising from AURKA inhibition, resulting in mitotic catastrophe and apoptosis. We found combined AURKA/WEE1 inhibition is potent in HNSCC xenografts, while well-tolerated in normal tissue and cells. We have extended this concept, identifying additional promising drug combinations between WEE1 and other G2/M regulatory kinases (PLK1 and CHK1). Notably, the limited number of cells surviving synthetic lethal treatment are characterized by aneuploidy and other defects suggesting they may have increased tumor mutation burden (TMB), express neoantigens, and upregulated inflammatory signaling associated with sensitivity to immune checkpoint inhibition. This project will take this observation directly to the clinic. We will conduct a pre-operative window phase I and expansion clinical trial of the late generation, high potency selective AURKA inhibitor LY3295668 with adavosertib combination, establishing pharmacodynamic proof of concept, identifying biomarkers for patient selection in future studies, and placing synergistic combinations in context of genomic alteration and treatment resistance. In Aim 1, we will evaluate mechanisms of combination lethality, and use single cell sequencing and Luminex profiling to query TMB, predict neoantigens, and measure inflammatory signaling. Aim 2 will query the effect of classes of common HNSCC TP53 and CDKN2A mutations, and cisplatin resistance, on response to a WEE1-AURKA inhibitor combination, using defined cell line models and patient derived xenografts (PDXs). In Aim 3, we will perform a pre-operative window trial to establish recommended phase 2 doses, determine activity and establish pharmacodynamic proof of concept, and to evaluate putative predictive biomarkers for response to combination WEE1/AURKA inhibition in HNSCC.

概括 HPV 阴性头颈鳞状细胞癌 (HNSCC) 通常会丢失 G1/S 细胞周期检查点， 大多数肿瘤的 TP53 发生突变，许多肿瘤还发生其他肿瘤抑制因子的突变，例如 CDKN2A。 这种肿瘤变得依赖于与 G2/M 相关的检查点来修复由 G2/M 引起的 DNA 损伤。 复制压力和其他基因组损伤。这种依赖性表明肿瘤选择性脆弱性 控制 G2/M 进展的合成致死策略。在广泛的初步数据中，我们表明 AZD1775/adavosertib 是 G2/M 检查点激酶 WEE1 的抑制剂，可有效致敏 TP53mut HNSCC 细胞系抑制极光 A 激酶 (AURKA)。 WEE1 诱导 CDK1 的抑制性 Y15 磷酸化， 阻断 M 期进入，从而减弱 AURKA 抑制的细胞毒性作用。 WEE1 抑制消除了这一点 停滞，加速具有高度破坏性纺锤体异常和其他缺陷的细胞进入有丝分裂 AURKA 抑制，导致有丝分裂灾难和细胞凋亡。我们发现 AURKA/WEE1 组合 抑制作用在 HNSCC 异种移植物中有效，同时在正常组织和细胞中具有良好的耐受性。我们已经扩展了这个 概念，确定 WEE1 和其他 G2/M 调节激酶之间其他有前景的药物组合 （PLK1 和 CHK1）。值得注意的是，在合成致死处理中幸存的细胞数量有限，其特点是 非整倍体和其他缺陷表明它们可能增加了肿瘤突变负担（TMB），表达 新抗原，以及与免疫检查点抑制敏感性相关的炎症信号上调。 该项目将把这一观察结果直接带到诊所。我们将进行术前窗口第一阶段和 新一代高效选择性 AURKA 抑制剂 LY3295668 的扩展临床试验 adavosertib 组合，建立药效学概念证明，识别患者的生物标志物 在未来的研究中进行选择，并在基因组改变和治疗的背景下进行协同组合 反抗。在目标 1 中，我们将评估组合致死机制，并使用单细胞测序和 Luminex 分析可查询 TMB、预测新抗原并测量炎症信号传导。目标 2 将查询 常见 HNSCC TP53 和 CDKN2A 突变类别以及顺铂耐药性对治疗反应的影响 WEE1-AURKA 抑制剂组合，使用确定的细胞系模型和患者来源的异种移植物 (PDX)。在 目标 3，我们将进行术前窗口试验，以确定推荐的 2 期剂量，确定活性 并建立药效学概念证明，并评估推定的反应预测生物标志物 联合抑制 HNSCC 中的 WEE1/AURKA。